XAF1在口腔鳞癌中的表达及对CAL27细胞生长和凋亡的影响

The expression of X-chromosome-linked inhibitior of apoptosis protein associated factor 1 in oral squamous cell carcinoma and its effect on cell growth and apoptosis of CAL27 cells

目的:研究X-染色体连锁凋亡抑制蛋白(X-chromosome-linked inhibitior of apoptosis protein,XIAP)相关因子1(XIAP associated factor 1,XAF1)在口腔鳞癌组织中的表达及临床意义,进一步观察过表达XAF1对口腔鳞癌CAL27细胞增殖、克隆形成和凋亡的影响。方法:应用免疫组织化学方法检测64例口腔鳞癌及14例正常口腔黏膜组织标本中XAF1蛋白的表达,分析XAF1表达水平与患者临床病理特征的相关性。CAL27细胞转染XAF1表达质粒,Real-time PCR检测转染24 h时XAF1 m RNA的表达;倒置显微镜观察细胞形态变化;CCK8和平板克隆实验检测过表达XAF1对细胞生长的影响;流式细胞仪和Western Blot检测细胞凋亡的影响。结果:XAF1蛋白在口腔鳞癌组织表达较正常口腔黏膜上皮组织降低。XAF1的表达与患者的TNM分期相关。过表达XAF1抑制口腔鳞状细胞癌CAL27细胞的增殖和克隆形成,凋亡细胞百分比和cleaved-PARP表达增加。结论:XAF1在口腔鳞癌组织中表达降低,可能与口腔鳞癌的进展相关。过表达XAF1抑制口腔鳞状细胞癌细胞的生长,促进细胞凋亡。

Objective： To explore the expression of X-chromosome-linked inhibitior of apoptosis protein associated factor 1（ XAF1） in oral squamous cell carcinoma（ OSCC） and investigate the effect of XAF1 overexpression on the proliferation,clone formation and apoptosis of CAL27 cells. Methods： The expression of XAF1 protein in 64 cases of oral squamous cell carcinoma and 14 normal oral mucosa tissues was detected by immunohistochemistry,and the correlation between the expression of XAF1 and the clinic opathological features of the patients was analyzed. CAL27 cells were transfected with XAF1 expression plasmid. Real-time PCR was used to detect the expression of XAF1 m RNA when transfected with 24 h. Inverted microscope was used to observe cell morphological changes. CCK8 and clones forming were used to detect the effect of XAF1 on cell growth. The effect of apoptosis was detected by flow cytometry and Western Blot. Results： The expression of XAF1 protein in OSCC was lower than that in normal oral mucosa. The expression of XAF1 was related to the TNM staging of the patients. Overexpression of XAF1 inhibited the proliferation and colony formation of CAL27 cells,and increased the percentage of apoptotic cells and cleaved-PARP. Conclusion： The expression of XAF1 in OSCC is decreased,which may be related to the progression of OSCC. Overexpression of XAF1 inhibits OSCC cell growth and promotes cell apoptosis.